Builing Traceability Systems Of Gannan Navel Orange Based On Metabolomics Technology

Gannan navel orange is a kind of orange,a rare specialty in Jiangxi Province.Gannan navel orange is popular with consumers for its color,flavor and taste.The chemical composition of Gannan navel orange is very complex,which is rich in a variety of active substances,these active substances have a variety of physiological activities,and have a significant effect on reducing the incidence of various diseases.Gannan navel orange has a complex composition,and the analysis of a single detector for traceability of its origin may be lacking,so it is necessary to use a combination of multiple analysis techniques.This paper analyzes the functional components and antioxidative activities of young fruit of southern Gannan navel orange,provides basis for the development and utilization of young fruit of Gannan navel orange,and uses SPME-GC-MS and UPLC-QTOF-MS/MS to analyze its volatile components.Qualitative analysis of water-soluble ingredients,and the use of metabolomics methods for traceability analysis and potentialbiomarkerexploration.Themain findingsareas follows:Chapter 1:expounded the chemical composition,its related physiological activity and recent research results of Gannan navel orange.Gannan navel orange has complex chemical components and rich nutrition.Its nutritional components include not only pectin,organic acids,oligosaccharides,various amino acids,but also bioactive substances that are beneficial to human health,such as polyphenols and carotenoids.The development of metabolomics at home and abroadandrelatedstatistical analysisarediscussed.Chapter 2:Total phenolic contents and total flavonoid contents of Gannan navel orange fruit of 6 different places of three origin of navel orange of Navel orange young fruit were firstly compared by Folin–Ciocalteu reagent method and sodium nitrite-aluminium chloride method.the antioxidant activities of these navel orange young fruit were evaluated by using2,2-diphenyl-1-picrylhydrazyl radicals(DPPH),ferric reducing antioxidant power(FRAP),and2,2′-azino-bis(3-ethylbenzthiozoline-6)-sulphonic acid(ABTS)methods.The correlation between antioxidant capacity and phenolic content was measured with Pearson correlation coefficient.Results showed that Total phenolic contents and total flavonoid contents navel orange young fruit varied from 11.02-17.14 mg/g,5.01-7.60 mg/g.navel orange in differentregions of young fruit polyphenol content,Xinfeng of navel orange young fruit total phenolic content and antioxidant capacity significantly less than Anyuan and Xunwu,total phenol and flavonoidsandantioxidantcapacity showedsignificantpositivecorrelation.Chapter 3: The study was to establish the fingerprints of Gannan navel orange from different growing areas and to make principal component analysis.The study carried out UPLC-QTOF-MS method to identify the common peaks of Gannan navel orange samples.The liquid-chromatographic separation conditions of this experiment were performed as followed: on a Agilent-C18 column(2.1 mm×100 mm,1.8 μm),using 0.1% formic acid aqueous solution(A)/methanol(B)as the mobile phase with a flow rate of 0.3 mL/min through gradient elution,column kept at 35℃,and detection wavelength was set at 254 nm,mass spectrometry thorough scan in positive ion modes.The fingerprint of Gannan navel orange was established by all above chromatograms,and was researched the similarity of the fingerprint.The common peaks acquired from fingerprints were used principal component analysis.Results showed that there were twenty-one common peaks in the fingerprints of fourteen batches of Gannan navel orange samples,and the similarity of samples was above 0.90 with their characteristic fingerprint.Therefore,this method could be used to distinguish the non-Gannan navel orange sample.Through principal component analysis,four principal components were obtained,whose cumulative contribution of variance accounted to 85.75% of the total variance.The samples can be classified into four groups.Through the analysis of the positive ion mass spectra information and the relevant reports,3 compounds of Common peak were identified.This method is accurate,reliable and well reproducible,and the established fingerprint combined with principal componentanalysis canbeusedfortheidentificationofthenavelorangeanditsorigin.Chapter 4: Volatile compounds of navel orange in Gannan and other areas by gas chromatography-mass spectrometry(GC-MS),and the metabolomics of navel orange in different areas was studied,which provided scientific basis for quality assurance of Gannan navel orange.The volatile components of navel orange were analyzed by GC-MS and identified by database matching with the relevant reports.The potential biomarkers were screened by the s-plot of orthogonal partial least squares discriminant analysis(OPLS-DA)and the variable importance in the projection value(VIP>1,p<0.05).The results showed that 66 volatile components were identified in Gannan navel orange,which were terpenes,aldehydes,alcohols,esters and ketones,the highest content was D-limonene.The five volatile components were(1S)-(+)-3-Carene,4-Carene,nonanal,(Z)-Caryophyllene,Eudesma-3,7(11)-diene,which can serve as potential biomarkers for the distinction between navel orange and other regions of the navelorange.Chapter 5: Determination of composition of Navel Orange in Gannan and Other Areas by UPLC-QTOF-MS.Study on the source of origin and potential biomarker of navel orange in south Jiangxi province based on metabonomics.The results showed that: using the secondary mass spectrometry to identify the Gannan navel orange species,according to the OPLS-DA modeling,the other origin of navel orange in Gannan navel orange is clearly distinguished,and preliminarily inferred 53 species the potential biomarker species and appraisal 18 species of Gannan navel orange from different producing areas.The establishment of a model based on metabolomics and the search for biomarkers provide a simple,rapid,and effective method for theidentificationoftheorigin of Gannan navelorange.